METHODS FOR GENERATING AMPLIFIED NUCLEIC ACID ARRAYS
First Claim
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1. A method of making an array, comprising(a) providing a genomic DNA sample comprising a plurality of annealed capture probes having different sequences that are complementary to different target regions of said genomic DNA sample;
- (b) sequentially treating said annealed capture probes with nucleotide analogs comprising reversible blocking groups under a first polymerase extension condition and then treating said annealed capture probes with nucleotide analogs comprising second blocking groups under a second condition, thereby producing a modified probe set comprising reversible blocking groups on a first plurality of said annealed capture probes and second blocking groups on a second plurality of said annealed capture probes,wherein said first polymerase extension condition has higher extension fidelity than said second polymerase extension condition;
(c) removing said reversible blocking groups from said modified probe set and then adding at least one nucleotide to deblocked probes of said modified probe set, thereby forming a plurality of different extension products comprising said target regions; and
(d) attaching said different extension products to an array.
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Abstract
The present invention relates to methods for generating an array of amplified nucleic acid sequences. The methods can utilize amplicons that form nucleic acid balls that can be arrayed on a solid support. The invention additionally provides methods for obtaining targeted nucleic acid sequences.
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Citations
15 Claims
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1. A method of making an array, comprising
(a) providing a genomic DNA sample comprising a plurality of annealed capture probes having different sequences that are complementary to different target regions of said genomic DNA sample; -
(b) sequentially treating said annealed capture probes with nucleotide analogs comprising reversible blocking groups under a first polymerase extension condition and then treating said annealed capture probes with nucleotide analogs comprising second blocking groups under a second condition, thereby producing a modified probe set comprising reversible blocking groups on a first plurality of said annealed capture probes and second blocking groups on a second plurality of said annealed capture probes, wherein said first polymerase extension condition has higher extension fidelity than said second polymerase extension condition; (c) removing said reversible blocking groups from said modified probe set and then adding at least one nucleotide to deblocked probes of said modified probe set, thereby forming a plurality of different extension products comprising said target regions; and (d) attaching said different extension products to an array. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15)
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Specification